Figure 7.

Pharmacological rescue of C84R and W174C by ML00253764. HEK293 cells stably expressing WT, C84R or W174C hMC4Rs or empty vector pcDNA3 were incubated with 10⁻⁵ M ML00253764 for 24 hrs. Cells were then stained with anti-myc antibody and visualized with a confocal microscope (A), or sorted with flow cytometry (B) to quantitatively measure cell surface expression. Cell surface expression of WT hMC4R treated with vehicle was set as 100%. In (A), the constructs transfected were WT (A1 and A2), C84R (A3 and A4), W174C (A5 and A6), and pcDNA3 (A7 and A8). A1, A3, A5, and A7 were treated with vehicle; A2, A4, A6 and A8 were treated with ML00253764. In (C), cells were stimulated with 10⁻⁶ M NDP-MSH and intracellular cAMP accumulation measured by RIA. Cyclic AMP generated by WT hMC4R treated with vehicle and stimulated with NDP-MSH was set as 100%. Star (⋆) indicates significant difference from corresponding vehicle treated controls (P < 0.05).